Ko Young-Joon, Choi Kang-Seuk, Nah Jin-Ju, Paton David J, Oem Jae-Ku, Wilsden Ginette, Kang Shien-Young, Jo Nam-In, Lee Joo-Ho, Kim Jae-Hong, Lee Hee-Woo, Park Jong-Myeong
National Veterinary Research and Quarantine Service, 480 Anyang-6 dong, Anyang, Kyong-gi 430-824, Korea.
Clin Diagn Lab Immunol. 2005 Aug;12(8):922-9. doi: 10.1128/CDLI.12.8.922-929.2005.
An inactivated SVDV antigen is used in current enzyme-linked immunosorbent assays (ELISAs) for the detection of antibodies to swine vesicular disease virus (SVDV). To develop a noninfectious recombinant alternative, we produced SVDV-like particles (VLPs) morphologically and antigenically resembling authentic SVDV particles by using a dual baculovirus recombinant, which expresses simultaneously the P1 and 3CD protein genes of SVDV under different promoters. Antigenic differences between recombinant VLPs and SVDV particles were not statistically significant in results obtained with a 5B7-ELISA kit, indicating that the VLPs could be used in the place of SVDV antigen in ELISA kits. We developed a blocking ELISA using the VLPs and SVDV-specific neutralizing monoclonal antibody 3H10 (VLP-ELISA) for detection of SVDV serum antibodies in pigs. The VLP-ELISA showed a high specificity of 99.9% when tested with pig sera that are negative for SVDV neutralization (n=1,041). When tested using sera (n=186) collected periodically from pigs (n=19) with experimental infection with each of three different strains of SVDV, the VLP-ELISA detected SVDV serum antibodies as early as 3 days postinfection and continued to detect the antibodies from all infected pigs until termination of the experiments (up to 121 days postinfection). This test performance was similar to that of the gold standard virus neutralization test and indicates that the VLP-ELISA is a highly specific and sensitive method for the detection of SVDV serum antibodies in pigs. This is the first report of the production and diagnostic application of recombinant VLPs of SVDV. Further potential uses of the VLPs are discussed.
目前用于检测猪水疱病病毒(SVDV)抗体的酶联免疫吸附测定(ELISA)中使用的是灭活的SVDV抗原。为了开发一种无感染性的重组替代物,我们通过使用双杆状病毒重组体产生了形态和抗原性上类似于天然SVDV颗粒的SVDV样颗粒(VLP),该重组体在不同启动子下同时表达SVDV的P1和3CD蛋白基因。在使用5B7 - ELISA试剂盒获得的结果中,重组VLP与SVDV颗粒之间的抗原差异无统计学意义,这表明VLP可用于替代ELISA试剂盒中的SVDV抗原。我们开发了一种使用VLP和SVDV特异性中和单克隆抗体3H10的阻断ELISA(VLP - ELISA)来检测猪血清中的SVDV抗体。用SVDV中和阴性的猪血清(n = 1,041)进行检测时,VLP - ELISA显示出99.9%的高特异性。使用从感染三种不同SVDV毒株之一的19头猪定期采集的血清(n = 186)进行检测时,VLP - ELISA最早在感染后3天就能检测到SVDV血清抗体,并且在实验结束前(感染后长达121天)一直能检测到所有感染猪的抗体。该检测性能与金标准病毒中和试验相似,表明VLP - ELISA是一种检测猪血清中SVDV抗体的高度特异性和灵敏的方法。这是关于SVDV重组VLP的生产和诊断应用的首次报道。还讨论了VLP的进一步潜在用途。